Method and apparatus for growing sprouts

ABSTRACT

Provided are methods for growing and shipping sprouts and microgreens in the same container, growing while in shipment using moisture provided in a water-absorbent layer, with optional added beneficials, and including methods for producing sprouts and microgreens for consumption, and for pharmaceutical/nutriceutical use, comprising growth of sprouts in retail-ready containers, the container comprising a moisture-retaining layer of agar media or the like providing water for growth and obviating the need for irrigation during sprout growth. In certain aspects, media is supplemented with beneficial organisms or additives such as probiotic microbes, vitamins (e.g., B12), cofactors, nutrients, and other items (e.g., phytochemicals, natural colors, and antioxidants) which promote the growth of the beneficial microbes on the product, and/or which become incorporated into the product. In certain aspects, added beneficial microorganisms are selected to compete/antagonize human pathogens such as  Listeria, Salmonella , enterohaemorrhagic  E. coli, Yersinia , and/or spoilage organisms (e.g.,  Erwinia, Pseudomonas  and  Xanthomonas ).

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/352,304, filed Apr. 16, 2014, which is a U.S. National Phase, under35 U.S.C. § 371, of International Patent Application No.PCT/US2012/060881, filed Oct. 18, 2012, which claims the benefit ofpriority to U.S. Provisional Patent Application No. 61/548,714, filedOct. 18, 2011, and U.S. Provisional Patent Application No. 61/654,571,filed Jun. 1, 2012, the disclosures of which are incorporated byreference herein in their entirety.

FIELD OF THE INVENTION

Particular aspects of the invention generally relate to sanitary methodsfor producing sprouts and microgreens for consumption (e.g., human, pet,and animal consumption), and in particular aspects, for pharmaceuticaland nutriceutical use, the methods providing for growth of sprouts in acontainer (e.g., retail ready/use ready container), and preferablywithout the need for further irrigation during growth.

BACKGROUND OF THE INVENTION

Sprouts, microgreens, shoots or crests (herein referred to as sprouts)are a food produced by exposing seeds to conditions which cause them togerminate and grow into tiny plants. This has previously been done byexposing seeds to moisture by various methods such as immersion orspray. Seeds which are germinated into edible small plant forms include,but are not limited to, those from alfalfa, clover, mung bean, radish,mustard, broccoli, flax, green peas, sunflower, corn, wheat, soybeans,and others.

Seeds that are used for sprouting purposes are obtained from sourcessuch as farm fields, and stored in places such as agriculturalwarehouses that may be contaminated with bacteria, fungi, or otherorganisms that can be harmful to health. For example, farm fields maybecome impacted with wildlife that carry microbial pathogens such as E.coli O157:H7, and salmonella, which can get spread to crops andultimately to the seeds. Seeds harvested from such fields, and sproutsgerminated from such seeds, may be contaminated (e.g., contaminated bysuch microbial pathogens).

Other organisms (e.g., spoilage organisms) with which sprouting seedsmay be contaminated may interfere with the quality of sprouts by, forexample, imparting to them a bad flavor or color, or by reducing shelflife.

Organisms harmful to the healthfulness and quality of sprouts may alsocome to be present with the seeds from sources other than seed harvestand transport. For example, water for soaking and spraying seeds andsprouts during washing and germination may be contaminated, as may bethe processing machinery surfaces and even process air used for aerationof sprouting chambers. Furthermore, workers working in a sproutmanufacturing plant can carry pathogens and transfer them to productsdirectly through handling of the products, and/or indirectly bycontaminating the production environment.

In the prior art, sprouts and microgreens are grown by methods whichinclude soaking, sanitizing the seeds, placing the seeds in trays, drumsor bags to allow for the growth to marketable size. Alternatively, seedsmay be placed in retail containers, with irrigation holes in the bottom,over a foam or an absorbent material such as cellulose and sprouted inthe same container in which they may ultimately be delivered toconsumers, placing the containers on trays and irrigating the growingsprouts from overhead or from the bottom tray with the seeds beingirrigated periodically via holes in the container. At the end of thegrowth cycle, the prior art containers are capped with a lid, labeledand sent to the market with open irrigation holes, which compromises thesanitary condition of the product and its package. Both of the foregoingprior art methods typically involve irrigation of the sprouts during allstages of their growth. For example, in one rotary drum method,sanitized seeds are grown for 4-6 days with constant rotation andirrigation of the drums, followed by washing of the sprouts and manualor automated packaging. At the end of the growth cycle, sprouts leavingthe drums typicalloyi contain about 10,000,000 to 100,000,000 cfu(colony forming units) of bacteria per gram. A subsequent wash andsanitation may reduce this load of bacteria by only a factor of 10 or100, but this reduction is rapidly undone by continued/more bacterialgrowth during subsequent storage.

With respect to prior art methods wherein the sprouts are grown in traysor containers (e.g., in the ultimate retail container), the trays andcontainers are typically placed on racks and are typically irrigated forthe length of the growth cycle by means of overhead irrigation, or bymeans of entry points in the bottom of the trays or containers, throughwhich the irrigation water may enter. The sprouts and microgreens,therefore, are exposed to contaminants that may enter with theirrigation water or with air that enters via the same pathway as theirrigation water, or via drainage holes that are provided for theirrigation water. Such holes for the entry and exit of irrigation waterallow microbial contamination to be introduced throughout the growthcycle.

There is a pronounced need in the art for methods/means for protectingthe healthfulness and quality of sprouts from pathogenic and spoilageorganisms, by containing the growth for most of the growth cycle in asanitary, closed, retail-ready container.

SUMMARY OF THE INVENTION

Aspects of the present invention provide methods for growing sprouts andmicrogreens that reduces the chance for growth of harmful microorganismsin the sprouts. In certain aspects, the methods allow for growing thesprouts for most of their growth cycle in a retail/consumer readypackage, for the first time allowing for growth to happen when theproducts are packaged, boxed and palletized, even during transportation.In certain aspects, the methods allow for full automation of sproutprocessing facilities, and/or allow for long-distance shipping ofsprouts, while they are growing. The methods also have the advantage ofproviding for increased shelf-life due to the sanitary condition of thegrowing environment and lack of post-process contamination.

In certain aspects, the present invention provides means for protectingthe healthfulness and quality of sprouts from pathogenic and spoilageorganisms, by containing the growth for most of the growth cycle in asanitary, closed, retail-ready container, thereby eliminating the needto irrigate the growing seeds and sprouts. In certain aspects, thecontainers used in the inventive method are closed to the outsideenvironment with the exception of allowing for exchange of air andgases. Additionally, in certain aspects, the inventive methodfacilitates the addition of a protective or beneficial microbiologicalflora that will compete with and resist any harmful organisms (e.g.,pathogen or spoilage organisms) that might otherwise be in, or beintroduced into the sprouts, with such beneficial microflora effectivelyacting as a protective immune system for the growing sprouts andmicrogreens. In certain embodiments, addition of beneficialmicroorganisms to the sprouts may be by direct application to the seedsand sprouts, and/or by adding such microorganisms to, for example, agarand/or other growth media in the container. In yet additional aspects,applied or added beneficial microbes may be selected which assist innitrogen fixation and/or promote plant growth. In yet further aspects,addition of beneficial probiotic organisms, vitamins, nutrients,cofactors, and the like (e.g., in each case to benefit the human andanimal consumers of the sprouts) is also provided and facilitated,because the inventive methods avoid the use of irrigation water on thesprouts, which irrigation water necessarily acts to wash such beneficialorganisms and additives away.

According to particular aspects, seeds or growing sprouts are sanitizedso as to reduce or eliminate any pathogens. Bad flora to be replacedinclude, but are not limited to, salmonella, Listeria, enterohemorrhagicEscherichia coli (EHEC), and spoilage organisms such as Psuedomonas.Then the seeds or sprouts are placed into a sanitized container with atleast one composition selected from the group consisting ofpasteurized/sterilized agar, agar-agar, seaweed, algae, algal extracts,plant derived polyesters such as cellulose, chitin, pectin, waterabsorbing natural fibers, foams, sponges and water absorbing polymers,or other water-absorbent compositions that can retain water and makesuch water available to the sprouts. Since the sprouts are thus providedwith a supply of water in the container which is sufficient to completetheir growth to marketable bulk, there is no need for additional waterto be introduced to the container by way of irrigation-therebyeliminating one pathway for contamination, and also permitting thesprouts to grow in containers while en route from the processingfacility to the ultimate consumer. In certain aspects, the media mayalso be provided with beneficial microorganisms which can, for example,produce natural antimicrobial substances that will assist in suppressingcontaminant organisms. Additionally, or alternatively, the media mayalso be provided with other microorganisms that produce vitamins (e.g.,B12), that add nutritional value to the sprout product. In certainaspects, added beneficial organisms may also compete with and thuscontrol the growth of harmful organisms. The seed or sprouts in thecontainer with the media layer may be promptly shipped from thepackaging facility, and then allowed to grow while in transit and whileon store shelves while waiting to be purchased. Vehicles used forshipping are preferably provided with temperature control to maintainproper temperatures for sprout growth and/or, later, storage. The sproutcontainers, not requiring additional input of water, may be sealed,except, preferably, for means such as a permeable plastic or celluloselayer, that permits gas exchange, while at the same time blocking entryof microorganisms and other contaminants.

In particular aspects, the present invention relates to a sanitarymethod for producing sprouts and microgreens for human, pet, and animalconsumption, and for pharmaceutical and nutriceutical use. Preferredembodiments of the invention provide for growth of sprouts in a sanitaryretail-ready or use-ready container without further need for irrigation.In one embodiment, the container comprises a bottom moisture-retaininglayer of agar or the like which provides water for growth, thusobviating the need for irrigation of the sprouts during the growthprocess. In certain aspects, the media can be supplemented withbeneficial additives such as probiotic microbes, vitamins, cofactors,nutrients, and other items which promote the growth of the beneficialmicrobes on the product or which become incorporated into the product.Preferably, beneficial additives may include vitamin B12,phytochemicals, natural colors, and antioxidants. Preferably, additivebeneficial microorganisms will be selected to provide antagonisticeffects on human pathogens such as Listeria, Salmonella,enterohaemorrhagic E. coli, Yersinia, and/or spoilage organisms (e.g.,Erwinia, Pseudomonas and Xanthomonas).

A particular embodiment of the invention comprises the steps of: a)placing a sanitized, non-toxic, preferably edible, water-absorbentmedium, which contains enough water for sprouts to grow to their fullintended size, into containers, which may be a retail-ready or use-readycontainer, with the water content of said medium being adequate tosupport growth of the sprouts without the need for irrigation; b)placing seeds or sprouts upon the medium; c) optionally providingbeneficial or probiotic organisms or nutritional factors; d) providingtemperature conditions to the containers suitable for promoting growthof sprouts in the container; e) optionally placing the containers intotransit in a temperature controlled shipping container or the like whilethe growth of step d) is taking place; and f) optionally, once sproutgrowth has completed to an adequate extent, shifting the temperature ofthe containers to a lower temperature suitable for storage and extendedshelf life. The temperature provided in growth step d) may preferably bein the range of 15-27° C. (60-80° F.), and the temperature provided instorage step f) may preferably be in the range of about 0-7° C. (32 to45° F.).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram of an exemplary container used in sprout productionaccording to certain aspects of the present invention.

FIG. 2 is a flowchart of a sprout production method production accordingto certain aspects of the present invention.

FIG. 3 is a detailed flowchart of the sprout production methodproduction of FIG. 2, according to certain aspects of the presentinvention.

DETAILED DESCRIPTION OF EXEMPLARY ASPECTS OF THE INVENTION

According to particular exemplary aspects, sprouts are grown upon alayer of media in a container such as a plastic “clam shell” or otherform of plastic or polymer based packages. Other usable containers are,for example, glass jars, ceramic jars, plastic tubs with lids, andgrowing trays with lids. The media may be or comprise any one of anumber of types, which have the common characteristic of beingwater-absorbent and structurally supportive of seeds or sprouts whichmay be placed in proximity. Agar and other types of gel are suitable,but the invention may be practiced with other water-absorbent materialssuch as cellulose fibers (e.g., at least one composition selected fromthe group consisting of pasteurized/sterilized agar, agar-agar, seaweed,algae, algal extracts, plant derived polyesters such as cellulose,chitin, pectin, water absorbing natural fibers, foams, sponges and waterabsorbing polymers, or other water-absorbent compositions that canretain water and make such water available to the sprouts).

Referring to FIG. 1, and embodiment of the inventive method for growingsprouts is as follows: container 101 which will hold the sprouts ispreferably first sanitized by application of a hydrogen peroxide,alcohol, ultraviolet light, steam, or other appropriate sanitizingagent.

Then layer 102, a sterile/pasteurized media containing appropriateamounts of water to complete the growth cycle, is placed into container101. If the chosen media is agar, this placement may be accomplished bypouring heated, liquid agar solution into the container and allowing itto cool. Layer 102 may be optionally inoculated with beneficialmicroorganisms as disclosed herein, and additionally there may be addednutrients to promote the growth of beneficial organisms. Other additivesto layer 102 may include antioxidants, vitamins, natural pigments,organic materials such as algae and seaweed, and plant nutrients, topromote the growth of the germination and growth of seeds and sprouts.

Seeds or sprouts are put into contact with water and allowed to absorbwater, and also sanitized. This step is important when a high density ofseeds is applied to the top of the gel layer, for two reasons: First, ifsufficient seeds are applied so that more than one layer of seeds comesto be present, then seeds in the upper layer will not be in directcontact with the gel, and will have no immediate source of water topromote germination. In this case, pre-soaked seeds which have absorbedwater may rely upon internal moisture to support germination and growthof an initial root down into the water-containing gel. Second, if a highdensity of seeds is placed upon the gel layer, then the gel proximate tothese seeds tends to become depleted of water as moisture is absorbedinto the many proximate seeds. The depleted gel is unable to supplysufficient water to allow adequate germination of the seeds. This isproblem is referred to herein as “gel top layer dry out.”

Preferably, the seeds are allowed to absorb at least about 40% of theirdry weight in water during the soaking process, and even morepreferably, 80% or more of their dry weight.

Sanitized seeds or sprouts 103 are placed in contact with layer 102 sothat they may grow with the benefit of the water contained in layer 102.Container 101 is preferably closed against microbial contamination, by,for example, placing a lid 104 on top of the container. Preferably, lid104 may comprise a portion or patch of plastic or other membranematerial that allows for exchange of gases, such as waste gases fromplant respiration. More preferably, the lid materials may further permitpassage of water vapor, so that excess condensation in the container maybe avoided. Optionally, a gas-exchange membrane may be provided at someother place on the container (other than the lid), or provision for gasexchange may be provided by means other than a membrane, such as bypermitting the lid to be slightly ajar, yet still placed so as toprevent entry of microbes, in the same manner of operation as a commonpetri dish lid. The point is that means be provided for the exchange ofgasses and water vapor which do not afford an entry point formicroorganisms. A key concept of this invention is maintaining layer 102and seeds 103 in a state of isolation from outside air and water, exceptsuch as might pass through a membrane or other means for excludingcontaminant microorganisms that may exist outside of container 101.

A preferred embodiment for practicing this invention is as follows, andreferring to FIG. 2. Seeds, for example alfalfa seeds, are obtained 200from a distributor. These seeds are sanitized 201, with or withoutpre-soaking, by soaking them in a solution comprising, for example, atleast one agent selected from the group consisting of bleach, hydrogenperoxide, organic acids, and/or other appropriate sanitizers with orwithout non-foaming surfactants. Alternatively, the seeds can, forexample, be flash-pasteurized by brief exposure to hot water. The seedsare then rinsed with water that may contain appropriate residualsanitizer levels. Although the seeds may be immediately placed upon thelayer of media and sealed into a container, it is best to soak the seedsso that they absorb water. Preferably, the seeds are allowed to absorbat least about 400% of their dry weight in water during the soakingprocess, and even more preferably, 800% or more of their dry weight.Seeds that have absorbed water tend to sprout and root better into themedium. Thus the seeds are soaked and drained 202. The moistened seedsmay also, optionally, be stockpiled in a refrigerator for a period oftime for later use.

After the optional refrigeration period, if any, the seeds are onceagain sanitized for an appropriate time and with appropriateconcentration of the sanitizing agents to reduce the bio-burden ofmicrobes attached to the seeds without impacting germination.Optionally, the seeds can then be pre-germinated by putting them into agermination chamber to germinate for 1-2 days, or can be directly placedinto contact with an agar layer 203 in the same container in which theywill be shipped. In the event that the seeds are pre-germinated, theycan be subjected to a third wash and sanitation protocol after whichthey can be placed into containers in step 203. The seeds germinate andproceed to grow 204 in the container. This growth may occur while thecontainer is in a holding area of a manufacturing facility and/or whileit is in transit 205 to the consumer and/or while the product isawaiting purchase by consumers on store shelves.

The layer of media on the bottom of the container may be of any suitablethickness, with the intent of the invention being best served by a layerwhich provides just enough volume of water to sustain sprout growthsufficient such that the sprouts come to fill the container. An optimumamount of agar would be such that the agar layer comes to be more orless completely absorbed after several days of sprout growth, and thecontainer appears to contain only sprouts.

It is important to emphasize that any gel media upon which the seeds areplaced must be suitable for human consumption (i.e., edible andpreferably palatable), since particles of the gel may be present withthe product. Preferably, this means that no non-edible germicide orother chemical may be added to the media.

The food products produced by the methods of this invention may beadditionally enhanced by various means. As described above in relationto FIG. 2, the seeds are preferably first sanitized 201 by washing in asanitizing solution such as bleach, for example, in a concentration ofno more than 20,000 ppm calcium or sodium hypochlorite. This soaking maybe for a period of time from, for example, 5 to 15 minutes or so, suchthat any bacteria are reduced in number but the seeds remain fertile.The subsequent sanitation of the hydrated seeds is done in much lowerdilutions of sanitizing agents to ensure that they are reducing themicrobial load without impacting germination. Thus, subsequent washingcan be done, for example, in 50-2000 ppm of calcium or sodiumhypochlorite. In certain aspects, it has been found that the seeds mustbe hydrated before being placed atop agar or other gel media if adequategermination rate and root penetration into the agar is to be obtainedwith the high densities of seeds required for adequate sproutproduction.

After the primary sanitizing treatment, and a wash to remove theresidue, the seeds may be preferably drained and refrigerated, andcultures of beneficial microorganisms may be added. These might includeyeasts or other microbes, which will provide vitamin B12, and commensualmicrobes that are antagonistic to human pathogens, plant pathogens, andspoilage organisms. The beneficial microorganisms added may consist ofone organism or comprise a cocktail of microbes blended for multipletasks, such as production of anti-microbial metabolites; production ofvitamins, cofactors, enzymes; and competition for nutrients withpathogens and/or spoilage organisms. The added beneficial microbialflora provides a replacement flora, given that the natural flora of theseeds is destroyed through aggressive sanitation, and is then replacedby beneficial organism(s) (e.g., an appropriate mix of mesophilic andpsychrotrophic/psychrophilic beneficial organisms). The beneficialorganisms may, for example, be spray inoculated onto the seed, such thatthey will tend to form bonds to the seeds and will adhere to them whenthe seeds are placed on the agar in step 203. In addition to addingbeneficials at this stage (e.g., to the sterilized seed), the agaritself may be inoculated when it is prepared or after it is placed intothe container.

“Beneficial” organisms include and in certain aspects are defined asharmless, edible microorganisms that can produce vitamins, enzymes, orcofactors that are healthful for humans, or which increase thenutritional value of sprouts and microgreens. Beneficial organisms incertain aspects may also inhibit pathogen and spoilage-organism growthby means of competition for space and nutrients and sometimes also bythe production of growth-inhibitory metabolites. Such beneficialorganisms may be selected to grow competitively at mesophilic growthtemperature range, which also tends to be the best range for sproutgermination and growth. Alternatively, such organisms may be selected togrow in the colder, psychrophilic range, thus affording best growthduring later shipping 205 and storage phases after sprout growth hasbeen accomplished and colder temperatures are employed to enhancestorage and shelf life.

“Beneficial microbes” or “beneficials” comprise living organisms such asbacteria, phages, yeast, viruses, fungi and the like which have theeffect of inhibiting growth of pathogenic or otherwise harmfulorganisms. The term beneficials used in this description also includesnatural antibiotic substances created and secreted by probioticorganisms, which substances tend to inhibit the growth of harmfulorganisms, for example, benign bacteriocins; and it also includes benignphages and viruses that attack pathogenic cells.

It is to be understood that the term beneficial microbes or beneficialsherein may also refer to mixtures of various probiotic species andvarious substances in addition to a single species or single substance.“Beneficials” used herein may also have the ability to deliverantagonistic properties against undesirable pathogens that mightotherwise come to be present in liquids used for sprout germination andgrowth.

One preferred group of microorganisms according to the present inventioncomprises lactic acid-producing microorganisms which inhibit pathogensby competing for substrate and generating a non-conducive acidicenvironment. The antagonistic properties of the lactic-acid producingmicroorganisms may additionally arise from other metabolites, likeenzymes (e.g., lactoperoxidases), pathogen-specific toxins, carbondioxide, peroxides or antibiotics, such as bacteriocins.

Beneficials may also include spore-forming organisms such as B.coagulans, Bacillus clausii, Bacillus pumilus, and other non-toxigenic,non-pathogenic Bacillus strains. These may have antimicrobial effectsagainst target organisms and have potential beneficial effects inhumans, for instance, as immunostimulants. These microorganisms have theability to create very quickly an environment that is not suitable forthe growth of pathogens. This is due to the rapid growth, high yield andreproducibility of such microorganisms in comparison to other lacticacid producing bacteria like Lactobacillus acidophilus. Spore-formingmicroorganisms are able to survive longer and reproduce themselves incomparison to non spore-forming microorganisms.

Exemplary suitable beneficial agents/organisms for use herein are lacticacid-producing microorganisms. Exemplary suitable lactic acid producingmicroorganisms for use herein are microorganisms that exhibitantagonistic properties against undesirable strains of microorganisms byreleasing amongst other metabolites, lactic acid.

Exemplary suitable lactic acid producing bacteria for use herein includeat least one selected from the genera group consisting of Lactobacillus(homofermentative members of the group), Lactococcus, Pedioccocus and/orLeuconostoc, and preferably the species Lactobacillus acidophilus,Lactobacillus curvatus, Lactobacillus plantarum, Lactobacillus jenseni,Lactobacillus casei, Lactobacillus fermentum, Lactococcus lactis,Pedioccocus acidilacti, Pedioccocus pentosaceus, Pedioccocus urinaeand/or Leuconostoc mesenteroides.

Aspects of the current invention allow for replacement of originalmicroorganisms found on the surface of a seed with a group offast-growing mesophilic organisms that can out-compete and potentiallykill residual target pathogens and spoilage organisms that have survivedthe sanitation process, and also allow for introducing a mixed cocktailof mesophilic and psychrotrophic beneficials either together or insequence, to allow the psychrotrophs to grow and protect the productduring refrigerated storage, with the mesophilic organisms havingpreviously protected the product during the germination and growthphases.

In a preferred embodiment, each beneficial strain used will becharacterized to ensure that it is void of virulence factors for humans,and is not antagonistic to other members of the cocktail or organisms tobe placed in contact with the seeds or agar.

When seeds and sprouts are in contact with moisture, an opportunity isprovided for pathogenic or other harmful organisms to take advantage ofthis moisture for purposes of survival and growth. According to thisinvention, beneficials are added to replace and out-compete the harmfulflora and prevent harmful growth from happening.

Beneficials may be brought into contact with sprouts in various ways,according to the apparatus and methods which are being employed togerminate and grow the sprouts. For this invention, beneficials may beadded after the seeds have been sanitized and their normal flora isdestroyed. Beneficials may also be added to the agar solution.

In particular aspects of the current invention, beneficials may be addedat any point in a sprouting process, from direct application to dryseeds to later application to the seeds as they rest upon the agarlayer, or by admixture with the agar layer as it is poured into thecontainer.

In a preferred embodiment of the invention, two groups of beneficialsare added to the media, with one group selected for optimum growth atmoderate temperatures (e.g., 15-27° C. (60-80° F.)), and the otherselected from growth at colder temperatures (e.g., 0-7° C. (32 to 45°F.)). The first group, known in the art as “mesophiles” will grow bestat the moderate, warm temperatures (e.g., 15-27° C. (60-80° F.)), whichthe seeds need to germinate and grow. The second group, known as“psycrophiles” or “psychrotrophs” grow best at colder temperatures(e.g., 0-7° C. (32 to 45° F.)), such as those at which the container ofsprouts will be stored after the germination and growth phases have beencompleted.

For example, if the best temperature for seed germination is 29-30° C.(85° F.), then the beneficials selected will be those which prefer togrow at or near this temperature. Even more preferably, if it is plannedthat the sprouts will be allowed, say, three days for growth, and thenbe placed in refrigerated storage at, say, 7° C. (45° F.), thenadditional beneficials, selected for this temperature, may also be addedto the media. The concept is that the first group of beneficials willprotect the sprouts while they are germinating, while the second groupwill multiply and take over this role when the container is cooled to 7°C. (45° F.), having remained more or less dormant at the highertemperatures.

A distinguishing feature of this invention is that it promotes promptshipping of sprout product from the manufacturing facility, as opposedto prior art systems wherein sprouts remain in the facility during theirentire growth phase, typically in a rotating drum being continuallyirrigated with water. In contrast, with the present invention, sproutsmay be shipped immediately after they are placed upon a layer of mediawithin the container.

Multiple containers may be placed upon, for example, a pallet, and thenthe pallet placed onto, for example, a truck or rail car for shipping.The truck or rail car is set up to provide the palletized containerswith heat or cooling to maintain optimum growth conditions for thesprouts during shipment. For example, if the sprout containers areplaced in a rail car, a heat pump unit on the rail car may be programmedto provide the best temperature for sprout growth. For example, if thebest temperature profile for a particular type of sprout is 21° C. (70°F.) for three days, with the sprouts then being placed into cold storageat 7-8° C. (45° F.), the rail car heat pump system would be programmedto provide this profile. Sprouts, for example, might be destined fordelivery to consumers in Boston, Mass., a five day rail trip from wherethey are packaged, in, for example, Spokane, Wash. In this case, seedswould be placed into containers with media and beneficials on day 1. Thecontainers would be sealed, placed in pallets, and loaded onto a railcar, where they would be kept at 21° C. (70° F.) for germination andgrowth on days 1, 2, and 3. On day 4, the sprouts will have completedtheir required growth, so, the rail car temperature would be reduced tostorage mode temperature, which could be 7-8° C. (45° F.). On days 4 and5, the sprout containers would proceed to Boston in the rail car at thecold storage temperature.

The foregoing is in contrast with the current art for delivering sproutsvia rail or truck, in which sprouts must reach desired size in themanufacturing facility before being loaded for transportation. In such asystem, the sprouts would not be growing during transit, but wouldmerely be in storage, with the transportation time being subtractivedirectly off the shelf life of the product. The end result in the priorart is sprouts that are several days less fresh being delivered toBoston.

A distinguishing feature of particular methods of the present inventionis that they allow shipment of sprouts with a variable temperatureprofile in the transportation vehicle, in which an initial growing phasetemperature is followed by a storage phase temperature. Thetransportation vehicle, which might also be a temperature-controlledshipping container, or a rail-car or truck, is provided with amultiphase time-temperature profile, having a first phase withtemperatures suitable for sprout growth, and a second phase withtemperature suitable for sprout storage. The temperature profile may,for example, be implemented by an automatic control system involving areal-time clock and a temperature controller, or it may be implementedby manual setting of the vehicle temperature controls in accordance withinstructions to the driver that are provided along with the sproutshipment.

This invention has various preferred aspects and/or advantages,including the following:

A. Growing in a retail ready package with no irrigation and no drainageholes. Particular aspects allow for initial input of the seeds and mediainto the ultimate retail package that will be delivered to the consumer,without the need for later input of water or nutrients by way ofirrigation. This means that the container need not have holes for theinput of irrigation water, or holes for the outlet of such water. Theabsence of these inputs and the holes they necessitate means that thecontainer may remain sealed against contamination, thus providing a moresanitary and healthful product, secure against contamination bypathogens.

B. Growing on a media that holds water, such as agar, cellulose,seaweed, algae, alginin, other complex polysaccharides or proteins, orpolymers. Agar is a suitable medium for the water-retaining layer uponwhich the seeds are placed. However, other water-retaining, food-safematrix materials are suitable for use with this invention, and thesematerials may include, for example, various forms of agar, cellulose,seaweed, algae, alginin, other complex polysaccharides or hydrophilicproteins, or polymers, sponges, textile fibers, and other suitablematerials that will be apparent, in view of the present disclosure, toone of ordinary skill in the art.

C. Growing container with micro-perforations or small holes that allowfor exchange of gases and water vapor without allowing for contaminantsto come in. The container in which the seeds and media are placed shouldresist the entry of microbial contaminants from the outside. This may bedone in a mechanical manner with a lid held by means, for example, ofgravitational force against the mouth of the container, in the manner ofa petri dish cover. Other arrangements may involve containers with smallholes and baffles which tend to block the entry of larger particles suchas airborne bacteria, while permitting the exchange of water vapor, airand respiratory gasses with the outside. Some plastics, textilematerials, and other materials permit gas and water vapor exchange whilealso blocking microbial contaminants. Medical gauze is an example ofsuch a material that might be used to provide a contamination-resistantvapor exchange port in a container otherwise made of imperviousmaterial. Various synthetic sheet materials are suitable for providinggas exchange while preventing microbial contamination. An example ofsuch material is 10 mil natural polyethylene film, as described inKordan, Applied Microbiology, Vol. 13, No. 5 (1965). Other suitablefilms might be those plastics used in blood banking operations thatpermit exchange of cellular gases, while preventing passage of liquidsand, of course, microorganisms. A discussion of suitable plasticmaterials is given in Christopher, Blood Banking and TransftsionMedicine, pp. 183-187, (Elsevier Health Sciences, 2007). FIG. 3 shows apatch of gas exchange material 310 incorporated into a container lid.

D. Seeding and growing with partial or complete growth in transit. Animportant advantage of particular aspects of the invention is provisionof the opportunity for the sprouts to grow while in transit (e.g.,without pathogen growth). After the media and seeds are placed into thecontainer, the container is sealed and needs no further inputs of wateror other materials. This means that the seeds may leave themanufacturing facility immediately and commence to grow while intransit, aboard a truck, railcar, or in a shipping container.Preferably, these modes of conveyance will provide temperature controltailored to the needs of the growing sprouts (i.e., warm temperatures(e.g., 15-27° C. (60-80° F.)) for the first few days in which growth iscommenced and then completed, and colder temperatures (e.g., 0-7° C. (32to 45° F.)) thereafter, to prolong shelf life). This is a significantadvance solving a major problem in the art of sprout growing because iteliminates the need for extensive plant floor space for sprout-growing,rotary bins or other machines, and, it also permits sprouts to beseveral days nearer to their ultimate destinations at the time whengrowth is completed and the clock starts to run on shelf life, andfurther permits a much higher level of safety with respect to potentialpathogens and spoilage organisms. With current technology, sprouts beginto age and burn shelf-life immediately upon packaging at the factory.However, with the current invention, sprouts may spend most or all oftheir transit time in the growth phase, and thus not enter theshelf-life use and decay phase until days later, providing major safetyand financial/commercial advantages.

E. Shipping in containers that hold the growth temperature and shift tocooling after desired growth is achieved. An important element ofparticular aspects of the invention is the use that is made oftemperature-controlled shipping means, such as refrigerated/heatedshipping containers, railcars, trucks, and the like. To take bestadvantage of this invention, filled sprout containers should be providedwith transportation or shelf-display at temperatures which best promotethe initial growth of the sprouts, and then lower temperatures that bestpromote storage, after optimum growth has been achieved. Thus,particular aspects of the invention afford and/or comprise the use oftransportation modes which provide temperature control suitable forspout growth in the initial hours and days until the sprouts reachmaturity, and then, preferably, which provide a switch to temperaturecontrol that is best suited to lengthening shelf-life, as in coldstorage. A computer controlled thermostat attached to a cooling/heatingunit on a shipping container is an example of such a mode, particularlyif the computer control also embodies a real-time clock than can changethe temperature when the sprouts have had enough time to grow, and thusneed to be maintained at a different, usually lower, temperature forstorage purposes. In cases where the transportation distance is notlarge, it may be that the sprouts may not have finished growth prior tothe time they reach the store where they are to be sold. In this case,the store itself may be provided with a temperature controlled displaycase, preferably programmed to switch over from growth-phase temperatureto storage-phase temperatures at the appropriate time. It is alsopreferable that each shipment of sprouts leaving the factory be providedwith remotely-readable data logging means that can be read by receivingstore display cases by means of a wireless data link, or the like, sothat the receiving store case may calculate the correct time to switchtemperature from the optimum growth phase temperature to the storagephase temperature.

F. A method of delivery of beneficial microbes, where the microflora ofthe seeds is destroyed through sanitation and replaced by the beneficialorganisms. An important element of particular aspects of the inventionis the opportunity it affords for the use of probiotic organisms toprotect sprouts from contamination during their growth and shippingphases. Prior art methods which involve irrigation prevent a lastingcontact to be made between probiotic organisms and the sprouts, becausethe sprayed-on irrigation water inherently washes off any beneficialsthat may have been previously applied. With this invention, however, thebeneficials may be incorporated into the media layer from the beginningor mixed in with the seeds that are applied to the media layer. Sincethe container is then sealed, there is no incoming irrigation or spraywater to wash away the beneficials. Moreover, the media may be providedwith nutrients suited to the growth of the probiotic organisms. It ispreferable that the seeds, container, and media be subjected tosanitation before use so that harmful microorganisms are eliminated orreduced. Then, a heavy dose (e.g., a dose sufficient for effectivecompetition/antagonism purposes relative to possible pathogens) ofbeneficial organisms is added to the container, so these are given agood start on growth without the need to compete with bad organisms.

G. A method for delivery of beneficial microbes. As discussed herein,beneficial microbes may be or include microbes that are antagonistic topathogens and spoilage organisms; microbes that are considered to bebeneficials for humans; and microbes that produce vitamins and cofactorssuch as vitamin B12, antioxidants, anti-carcinogens, anti-teratogens,immunostimulants, and beneficial oils. These microbes may includemesophilic microbes added at an early stage in the method to sanitizedseeds and early-stage sprouts to allow for competition against pathogensand spoilage organisms. A second group of psychrotrophic orpsychrophilic beneficial microbes may be simultaneously added, or areadded at the end of the growth cycle. The members of this second groupgrow at lower temperatures and thus offer protection to the product atlow temperatures which are used to extend shelf-life of the product,subsequent to the sprouts reaching marketable size. An important aspectof particular aspects of the invention is the concept of two-phasegrowth of the product: A first phase for germination and development ofthe sprouts to marketable size, which is accomplished, generally at warmtemperatures conducive to plant growth, and, a second phase, in whichthe sprouts are subjected to different, usually colder, temperature, sothat the shelf-life of the mature product may be improved. Thistwo-temperature-phase aspect is not only important for transportationarrangements, as described above, but it is also important from thestandpoint of selecting the best probiotic organisms. Various probioticorganisms may have different temperature preferences for their ownmultiplication and growth. Some organisms are “mesophiles” which likewarmer temperatures, such as room temperature, or perhaps higher, suchas 4-5° C. to 7-8° C. (40 to 45° F.). These organisms are best suited toprotect the sprout product during the germination and growth and phase,when the sprouts also develop best with warm temperatures. However, whenthe sprouts reach market size, and need to go into the storage, colderenvironment to preserve shelf life, then the mesophilic beneficials maynot remain effective. At this point, cold-loving, psychrophilicprobiotic organisms are needed. These organisms can be provided byincluding them in the initial input of beneficials into the media ofthis invention. These organisms will remain dormant during the initialwarm-phase of the product, but, when storage-phase temperatures areimposed, they will come to life and multiply, to provide probioticprotection and benefits during storage as well. Preferred aspectsprovide a mixture of probiotic organisms, containing bothmesophilic/warm-loving organisms and psychrophilic/cold-loving organismsin the initial input of beneficials.

H. Another beneficial aspect of this invention is the ability to imposesafety and quality testing at various stages of the method. In certainaspects, the invention comprises sprout and micro-green production inwhich the safety of the product is assessed with one or more of thefollowing steps:

-   -   a. Each batch of seed is subjected to statistically significant        sampling and testing for multiple pathogens including        Salmonella, Listeria, and EHEC;    -   b. After 1-2 days of germination, germinated sprouts are        subjected to secondary testing for the same organisms;    -   c. A finished product testing program, employing a statistically        significant sampling plan is employed to test the finished        product before release into the market; and    -   d. A method of conducting finished product sampling where        retail-ready containers are seeded, samples are taken and placed        in a larger container with the media that will be subjected to        growth at the same or preferably higher temperature to expedite        growth and then tested before the release of the lot.

I. Addition of Microbe-Growth-Promoting Nutrients to the Agar to promotegrowth of the probiotic organisms. To encourage growth of the probioticorganisms in the media layer of embodiments of the invention, nutrientsthat enhance the growth of probiotic organisms may be added, such asthose found in DIFCO™ MRS media, the formula for which is as follows:

DIFCO ™ Lactobacilli Agar AOAC Approximate Formula* Per Liter PeptonizedMilk 15.0 g Yeast Extract 5.0 g Dextrose 10.0 g Tomato Juice (from 100mL) 5.0 g Monopotassium Phosphate 2.0 g Polysorbate 80 10 g Agar 10.0 gApplicant has also been found that adding a source of nitrogen andminerals can promote growth. For example, enhanced growth was observedwhen a sheet of dried kelp was added beneath the agar layer.

To provide an adequate number of sprouts in a retail package, it ispreferable to place sufficient sprouts upon the agar layer in thecontainer such that at least a portion of that layer is covered with adouble layer of sprouts. It is more preferable that said layer betotally covered with two layers of sprouts, and, in some cases threelayers of sprouts. To provide a source of oxygen for sprout development,it has been found beneficial to add to the water absorbent layer amicroorganism that produces hydrogen peroxide.

In the following claims, the term “sprouts” includes sprouts, shoots,and microgreens. Microgreens are defined in the food trade as greens,lettuces and herbs which are harvested when quite young, generally whenthey are approximately an inch tall. The term sprout also includesnewly-germinated seeds and also small plants which are created frommethods of tissue culture or asexual production which do not involveseed. In particular embodiments of the invention, sprouts also comprisesoaked seeds used for sprout generation, which soaked seeds are placedon the water-absorbent layer in the methods of the invention.

EXAMPLE 1 A No. 1305 100 mL PYREX Glass Container was Prepared withabout 10 ml of 0.5% Agar Solution

A No. 1305 100 mL PYREX glass container was prepared with about 10 ml of0.5% (wt %) agar solution. This concentration was chosen as optimumafter some experimentation, as it was found that 0.7% (wt %) solution,normal for bacteriological use, was too rigid for easy root penetrationand sprout embedding, whereas a 0.4% (wt %) solution provided a gelwhich was too soft, such that some of the larger seeds or sprouts sunkto the bottom of the agar layer. The preferable consistency for thelayer is to be as soft as possible yet still rigid enough to support theseeds on the top of the layer (e.g., between 0.4% (wt %) and 0.7% (wt%), preferably between 0.45% (wt %) and 0.6% (wt %), or 0.45% (wt %) and0.55% (wt %)). A stock 0.5% (wt %) agar solution was prepared with 0.5 gof agar power in 100 ml of water. Rather than agar, other water-holdingmaterials may be substituted, for example, alginate, complex naturalpolysaccharides, cotton fiber, wood fiber, paper, protein gels, and thelike. Any substance which can hold water adequate for growth of theseeds is a candidate, provided it is not easily biodegradable ormetabolized by the microbial flora of the seeds or the introducedbeneficial microbial flora. Agar is a preferred substance because it isfood grade and is a natural product of seaweed. Agar is also preferablebecause it becomes essentially invisible as the contained water isabsorbed into the sprouts, such that after a few days of growth, layer102 (see FIG. 1) virtually disappears and only a mass of roots is seenin its place.

EXAMPLE 2 Sprout Seeds are Sterilized, Rinsed, Soaked, Placed on MediaLayer in Container Sealed with Cap Permitting Gas Exchange, andProviding for Growth in Container, Even During Shipment and DuringDisplay on Store Shelves

FIG. 2 merely illustrates one embodiment of the invention, which couldbe implemented with various seeds, washing methods, containers, andprobiotic additions. FIG. 3 depicts another embodiment of the inventivemethods for growing sprouts of FIG. 2. Sprout seeds 300 are placed intobag 301 and dipped into container 303 containing a bleach solution 304.The bleach solution residue is removed from the seeds by rinsing the bagwith water. Then the seeds are placed into soaking solution 306contained in container 305, which optionally contains beneficials. Aftera suitable period of soaking, which typically may be from six toforty-eight hours, the seeds are either close to germination or havealready germinated. At this point, the seeds 300 are placed into sterilecontainer 307 upon media layer 308 at the bottom of the container, andthe container is sealed with cap 309. The cap may be held in place bygravity, providing a physical barrier against microbial infiltration,while also permitting gas exchange, in the same manner as a petri dishcover. Optionally, the container cap could be sealed tightly, but beprovided with permeable membrane portion or patch 310, which is a patchof gas permeable plastic or other suitable material, which permits gasexchange to occur while not affording passage to objects the size ofbacterial cells (e.g., 1 to 2 microns). According to particular aspects,a gortex-type material (polytetrafluoroethylene, the chemicalconstituent of Teflon™, with a micro-structure characterized by nodesinterconnected by fibrils) may be used.

A great advantage of this approach is that sprouts are allowed to growin container 307, even during shipment 205 and during display on storeshelves. This means that shipment from the manufacturing facility maytake place as soon as sprouts are placed in the container, without theneed for several days of additional growth, which in the prior art,would occur in drums or like apparatus at the manufacturing facility.This greatly reduces requirements for facility floor space, and it alsomeans that sprouts will be available for consumption immediately aftercompleting growth during the shipping phase. The results in addedproduct shelf-life and fresher, safer sprouts available for theconsumer.

It should be noted that the seeds and sprouts of this inventionpreferably form a dense growth of plants upon the surface of the media,the roots of which eventually absorb the water from media layer 308,causing that layer to shrink or disappear. At this point, the volume ofcontainer 307 consists almost completely of sprouts.

Agar-like materials or other water-holding materials could besubstituted for agar in the practice of the methods of the invention.The role of agar (or the other suitable material) is to hold moisture ina confined format and provide a structural support upon which thesprouts may rest. Substances such as animal protein gelatin, which alsoholds water in a structural matrix, could also be used. One might alsouse a layer of sponge or textile or other water-absorbent material toprovide support and water to the seeds. This is less preferable sincethe consumer would need to remove the bottom layer and discard it uponopening the product for use.

1. A method for growing sprouts during shipping, comprising the stepsof: placing, prior to shipping, seed or germinated seeds in contact witha water supplying material layer in a sanitary, closed, retail-readycontainer, such that the water supplying material layer is covered bymore than one layer of the seeds or germinated seeds so that the seedsin the upper layer of the seeds or germinated seeds are not in directcontact with the water-supplying material layer; and shipping thecontainer, wherein during shipment the container is configured toprovide for exchange of gases and water vapor while precluding entry ofcontaminants, wherein the water-supplying material layer providessufficient water to provide for growth of the sprouts to marketable bulkor size without the need for additional water input or opening thecontainer, and wherein the seeds or germinated seeds, prior to placingin contact with the layer of water-supplying material, are derived fromhydrated seeds with internal moisture sufficient to support germinationof the seeds in the upper layer of the seeds or germinated seeds thatare not in direct contact with the water-supplying material layer, andprevent dry out of the top of the water-supplying material layer indirect proximate contact with the seeds or germinating seeds.
 2. Themethod of claim 1, wherein said water supplying material layer comprisesone or more selected from the group consisting of agar, agar-agar,seaweed, algae, algal extracts, alginate, cellulose, chitin, pectin,complex natural polysaccharides, sponges, water absorbing polymersfiber, paper, and protein gels.
 3. The method of claim 1, wherein thewater-supplying material is divided into pieces so as to afford bettercontact with the sprouts, seeds of germinated seeds.
 4. The method ofclaim 1, wherein the water-supplying material supplies sufficient waterto grow the sprouts to marketable size, without the need for additionalwater input, to provide for avoiding microbial contamination.
 5. Themethod of claim 1, wherein the water supplying material layer suppliessufficient water to provide for growth of the sprouts without the needfor additional water input during shipment or prior to sale.
 6. Themethod of claim 1, wherein the seeds or germinated seeds, prior toplacement, are soaked to absorb at least about 40% of their dry weightin water during the soaking process.
 7. The method of claim 1, whereinthe sprouts, seeds or germinated seeds, prior to placement, are derivedfrom seeds soaked in the presence of a beneficial microbial organism. 8.The method of claim 1, wherein the container is sealed against theintroduction of liquid water.
 9. The method of claim 1, wherein thecontainer is a retail-ready container sealed against the introduction ofliquid water and microbial contaminants.
 10. The method of claim 1,comprising the additional step after achieving adequate growth, ofmaintaining the container at a different temperature appropriate forsprout storage, wherein the storage temperature is different than thesprout growth temperature.
 11. The method of claim 1, wherein the watersupplying material layer comprises at least one additive selected fromthe group consisting of: beneficial microorganisms, probioticmicroorganisms, vitamins, vitamin B12, cofactors, nutrients,phytochemicals, natural colors, antioxidants, plant nutrients, and humannutrients.
 12. The method of claim 11, wherein the selected additivecomprises a microorganism capable of producing hydrogen peroxide. 13.The method of claim 11, wherein the beneficial microorganism is at leastone selected from the group consisting of mesophilic microbes, andpsychrotrophic or psychrophilic microbes.
 14. The method of claim 13,wherein the added beneficial microorganisms comprise both mesophilicmicrobes and psychrotrophic or psychrophilic microbes.
 15. The method ofclaim 14, wherein the seeds or germinated seeds are sanitized, whereinthe mesophilic microbe is added at an early stage in the method to thesanitized seeds or the germinated seeds, and wherein the psychrotrophicor psychrophilic microbe is added at a later stage, in each case insufficient amounts to allow for competition against pathogens andspoilage organisms at the respective stages.
 16. The method of claim 1,further comprising growing the sprouts to a marketable bulk or size. 17.The method of claim 1, further comprising shipping the container undertemperature conditions such that the sprouts continue to grow duringshipping.
 18. The method of claim 1, wherein the seeds are hydrated andinoculated with beneficial organisms and/or probiotics, and refrigerateduntil being placed in contact with the water-containing material. 19.The method of claim 1, wherein, after placing the seeds or germinatedseeds in contact with the layer of water-supplying material in thecontainer, the container is packed and shipped, and growth to marketablebulk or size is achieved in transit.
 20. The method of claim 2, whereinthe water-supplying material layer comprises agar, agar-agar, or water.21. The method of claim 6, wherein the seeds or germinated seeds, priorto placement, are soaked to absorb at least about 80% of their dryweight in water during the soaking process.
 22. The method of claim 1,wherein growth of the sprouts to marketable bulk or size comprisesgrowth at a temperature in the range of 40° F. to 45° F.
 23. The methodof claim 22, wherein growth of the sprouts to marketable bulk or sizecomprises growth at 40° F.
 24. The method of claim 10, wherein thedifferent temperature appropriate for sprout storage is lower than thesprout growth temperature.